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Two-color flow cytometric analysis of CD4+CD25+ cells and subdivision of these cells tool into CD4+CD25high Treg cells and CD4+CD25low cell subsets. First, lymphocytes gated in FSC (forward light scatter)–SCC (side light scatter) dot plot and then CD4+CD25- and CD4+CD25+ (high and low) gated in CD4-FITC intensity versus CD25-PE intensity.
